Biosynthesis of poly-gamma-glutamic acid in plants: transient expression of poly-gamma-glutamate synthetase complex in tobacco leaves

Transient expression of genes coding for the poly-gamma-glutamate (gammaPGA) synthetase system (pgs) was investigated in tobacco plants. Three genes of the pgs, pgsA, pgsB and pgsC, were separately placed under the control of the CaMV 35S promoter and introduced into tobacco leaves via Agrobacterium infection. Synthesized gammaPGA in plant tissues was detected immunologically with mouse anti-gammaPGA antiserum which specifically reacts with gammaPGA on a nitrocellulose membrane. Confirmation of gammaPGA biosynthesis in the transient expression analysis in tobacco tissue indicates that subunits of pgs complex were expressed and reassembled in a functional form.     

Penicillium camemberti and Penicillium roqueforti enhance the growth and survival of Shiga toxin-producing Escherichia coli O157 under mild acidic conditions

The effects of secondary starter molds of common mold-ripened cheeses on the Shiga toxin-producing Escherichia coli (STEC) O157 were assessed in 3 model systems. In the 1st model, 8 STEC O157 strains were incubated in the spent culture of Penicillium camemberti or Penicillium roqueforti under mild acidic conditions at 25 °C. In the spent cultures of the mold at pH 4.8 to 5.0, the lag times of STEC O157 growth were significantly shorter than those observed in fresh medium. Analyses of the spent culture of P. camemberti showed that the causative agents of the growth enhancement were produced by the mold in response to an acidic environment and were not fully inactivated in heat treatment. In the 2nd model, P. camemberti and STEC O157 were cocultured in acidified milk at 25 °C. The population of STEC O157 reached 10(8) CFU/mL in the presence of the mold, whereas the population steadily declined in the absence of the mold. Although this growth enhancement was partially attributable to alkalization by the mold, it was observed even when the pH of this model was stabilized. In the 3rd model, 2 STEC O157 strains were incubated in the spent cultures of molds at pH 4.5 at 10 °C. In the spent culture, proportions of injured cells were significantly lower and D values were significantly higher than those in control, except one STEC O157 strain in the spent culture of P. camemberti. These results showed that the molds could enhance the growth and survival of STEC O157 by changing the environment. Practical Application: This study demonstrated that molds in foods can improve the growth and survival of the Shiga toxin-producing Escherichia coli O157. Because microbial interactions are ubiquitous in food, our results provide an important insight for understanding the behavior of microorganisms in food.     

Evidence that activation of Src family kinase is not required for fertilization-associated [Ca2+]i oscillations in mouse eggs

Recent evidence in marine invertebrate, frog, and zebrafish eggs suggests the involvement of a Src family kinase (SFK) in fertilization-induced Ca2+ release. In the present study, we have investigated whether activation of an SFK is required for initiation of intracellular Ca2+ ([Ca2+]i) oscillations in mouse fertilization. We detected a Hck-like protein and tyrosine-phosphorylated proteins in soluble and insoluble sperm fractions, respectively. However, the presence of these proteins did not correspond to the active fractions of porcine sperm extracts (pSE). Moreover, [Ca2+]i oscillations induced by pSE in mouse eggs were unaltered by pre-incubation of pSE with specific SFK inhibitors such as 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazol[3,4-d]-pyrimidine (PP2) or lavendustin A, despite the fact that the inhibitors were shown to be active both in vivo and in vitro. Another SFK inhibitor, peptide A, blocked oscillations when incubated with pSE prior to injection into eggs, but this inhibition required more than ten times the concentration reportedly required to inhibit SFK activity. In addition, pre-injection or pre-incubation of eggs with these inhibitors did not affect the ability of pSE to trigger [Ca2+]i oscillations in mouse eggs. Microinjection of a recombinant c-Src protein or mRNAs encoding constitutively active Src proteins did not induce [Ca2+]i release. Finally, when sperm and eggs, both of which were pre-treated with PP2, were fertilized, [Ca2+]i oscillations occurred normally. We can therefore conclude that activation of an SFK is neither necessary nor sufficient for triggering fertilization-induced [Ca2+]i oscillations.     

Dependence of Diamond Transition on Microtexture in Graphite Starting Materials under Shock Compression

The effect of microtexture on diamond transition was examined for graphite starting materials under shock compressions of 50 to 60 GPa and 80 to 90 GPa. Each of the starting materials used in the present study possessed a fully homogeneous microtexture. To distinguish the effect of microtexture from that of other experimental parameters, the shock conditions were standardized for all specimens tested. Three graphite materials—a glassy carbon, a carbon black, and a natural graphite—were selected and shock compressed using a quenching technique to generate conditions common to all samples. Detailed characterization by transmission electron microscopy and electron energy-loss spectroscopy revealed a clear tendency: The lower the crystallinity and crystallite size of the starting graphite, the more easily the graphite transformed to diamond when the transition mechanism was reconstructive.     

Difference in environmental degradability between poly(ethylene succinate) and poly(3-hydroxybutyrate)

A chemosynthetic aliphatic polyester, poly(ethylene succinate) (PESu), was degraded by a poly(3-hydroxybutyrate) (P(3HB)) depolymerase in vitro. While P(3HB) exhibited good biodegradability in all environments, PESu hardly underwent biodegradation in a marine environment. To understand the difference in environmental degradability between PESu and P(3HB), we investigated the distribution of P(3HB)- and PESu-degrading microbes in various environments. PESu-degrading microbes were never detected in marine environments. PESu-degrading bacteria isolated from various environments in this study belonged to the phyla Firmicutes and Proteobacteria. Most PESu-degrading bacterial isolates could not degrade P(3HB), suggesting that PESu was not degraded by P(3HB) depolymerase in actual environments. In addition, all bacterial isolates that were screened for P(3HB) degrading activity from various environments in this study did not degrade PESu, suggesting that PESu does not induce P(3HB) depolymerase in their bacteria and P(3HB)-degrading bacteria are not involved in biodegradation of PESu in actual environments. Taken together, these results could be related with the low biodegradability of PESu in marine environments.     

Effect of Substituents on Dielectric β-Relaxation in Cellulose

The dielectric characteristics for some cellulose derivatives, namely chlorodeoxycellulose (Cell-Cl; degree of substitution of chlorine, DS_Cl=0·87), bromodeoxycellulose (Cell-Br; DS_Br=0·92) and thiocyanatodeoxycellulose (Cell-SCN; DS_SCN=0·88), all substituted only at C-6, together with those of regenerated cellulose, have been investigated in the temperature range -60 to 120°C, and in the frequency range 0·2–100kHz. Only one relaxation process, designated as β, was identified within the frequency and temperature ranges studied. The activation energy of this relaxation increases in the order Cell-Cl<Cell-Br<Cell-SCN, suggesting that the bulkiness of the substituent was the determining factor of the activation energy. The characteristic dielectric parameters, namely polarization magnitude (Δε) and shape parameter (α or β-), were obtained by the analysis of absorption bands and are discussed in relation to the substituent effect. © of SCI.     

Cooperative behavior of various agents in dynamic environment

The multi-agent model is a model in which agents with limited ability cooperate each other to accomplish a goal. In this paper, we introduce a multi-agent model in which agents are created to imitate real ants. There are two different type of agents, each type of which has a particular task. We designed agents to communicate each other by using pheromone considering noise. On this model, we observed cooperative behavior of agents and evaluated conditions to make an order of behavior in the model.     

Structures and electronic properties of surface/edges of nanodiamond and nanographite

The structure, electronic and magnetic properties of nanodiamond and nanographite/nanographene are investigated. Detonation nanodiamond particles that are covered with amorphous graphitic composites are hydrothermally treated to remove the graphitic surface composites and to terminate the surface carbon atoms with hydrogen. The number of localized spins of dangling bonds and the hydrogen concentration increase upon the increase in the hydrothermal treatment time up to 40 h. Above 40 h, both drop discontinuously, a surface structural reconstruction was suggested. The creation of dangling bonds and an incomplete hydrogenation of the surface carbon atoms destabilize the surface, resulting in the structural reconstruction. Nanodiamond particles are thermally converted to nanographite/nanographene. A single nanographene sheet is successfully prepared by heat-treating nanodiamond particles. The edge of graphene sheet with its edge carbon atoms being hydrogen-terminated is investigated by UHV-STM/STS. Zigzag edges are found to have non-bonding π-state of edge origin, in good agreement with theoretical prediction.